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Preparative Mini-Slab Gel Continuous Elution Electrophoresis: Application for the Separation of Two Isoforms of Rat Hepatocyte Cell Adhesion Molecule, Cell-CAM 105, and Its Associated Proteins

✍ Scribed by Y.P. Lim; H. Callanan; S.H. Lin; N.L. Thompson; D.C. Hixson


Publisher
Elsevier Science
Year
1993
Tongue
English
Weight
875 KB
Volume
214
Category
Article
ISSN
0003-2697

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✦ Synopsis


A simple method for performing continuous elution electrophoresis using a mini-slab-gel system has been developed. The high resolving power of this method provides a means to separate complex mixtures of proteins and recover individual peptides in high yields. Recoveries between 89 and (99 %) were achieved using a single percentage gel to separate a mixture of standard proteins radiolabeled with ({ }^{125}) I. Peptide fragments between 2.5 and (60 \mathrm{kDa}) generated by (\mathrm{CNBr}) digestion of albumin were isolated within (150 \mathrm{~min}). Application of continuous elution electrophoresis to the purification of proteins associated with the rat hepatocyte cell adhesion molecule, cell-CAM 105 , resulted in complete separation of the two predominant isoforms and the purification of three immunologically unrelated proteins that were coisolated during immunoaffinity chromatography. 厄 1993 Academic Press, Inc.